The antifibrotic potential of a sustained release formulation of a PDGFβ-receptor targeted rho kinase inhibitor

F van Dijk; N Teekamp; E Post; D Schuppan; Y O Kim; J Zuidema; R Steendam; Matthias H M Klose; Samuel M Meier-Menches; A Casini; P L Horvatovich; N J Sijbrandi; H W Frijlink; W L J Hinrichs; K Poelstra; L Beljaars; P Olinga

doi:10.1016/j.jconrel.2018.12.039

The antifibrotic potential of a sustained release formulation of a PDGFβ-receptor targeted rho kinase inhibitor

J Control Release. 2019 Feb 28:296:250-257. doi: 10.1016/j.jconrel.2018.12.039. Epub 2019 Jan 22.

Authors

F van Dijk¹, N Teekamp², E Post³, D Schuppan⁴, Y O Kim⁵, J Zuidema⁶, R Steendam⁶, Matthias H M Klose⁷, Samuel M Meier-Menches⁸, A Casini⁹, P L Horvatovich¹⁰, N J Sijbrandi¹¹, H W Frijlink², W L J Hinrichs², K Poelstra³, L Beljaars³, P Olinga¹²

Affiliations

¹ Groningen Research Institute of Pharmacy, Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Groningen, The Netherlands; Groningen Research Institute of Pharmacy, Department of Pharmacokinetics, Toxicology and Targeting, University of Groningen, Groningen, The Netherlands.
² Groningen Research Institute of Pharmacy, Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Groningen, The Netherlands.
³ Groningen Research Institute of Pharmacy, Department of Pharmacokinetics, Toxicology and Targeting, University of Groningen, Groningen, The Netherlands.
⁴ Institute of Translational Immunology and Research Center for Immune Therapy, University Medical Center, Johannes Gutenberg University, Mainz, Germany; Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA.
⁵ Institute of Translational Immunology and Research Center for Immune Therapy, University Medical Center, Johannes Gutenberg University, Mainz, Germany.
⁶ InnoCore Pharmaceuticals, Groningen, The Netherlands.
⁷ Department of Analytical Chemistry, University of Vienna, Waehringer Straße 38, 1090 Vienna, Austria.
⁸ School of Chemistry, Cardiff University, Park Place, CF10 3AT, Cardiff, UK; Department of Analytical Chemistry, University of Vienna, Waehringer Straße 38, 1090 Vienna, Austria.
⁹ School of Chemistry, Cardiff University, Park Place, CF10 3AT, Cardiff, UK.
¹⁰ Groningen Research Institute of Pharmacy, Department of Analytical Biochemistry, University of Groningen, Groningen, the Netherlands.
¹¹ LinXis BV, Amsterdam, The Netherlands.
¹² Groningen Research Institute of Pharmacy, Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Groningen, The Netherlands. Electronic address: p.olinga@rug.nl.

PMID: 30682444
DOI: 10.1016/j.jconrel.2018.12.039

Abstract

Rho kinase activity in hepatic stellate cells (HSCs) is associated with activation, transformation and contraction of these cells, leading to extracellular matrix production and portal hypertension in liver cirrhosis. Inhibition of rho kinase activity can reduce these activities, but may also lead to side effects, for instance systemic hypotension. This can be circumvented by liver-specific delivery of a rho kinase inhibitor to effector cells. Therefore, we targeted the rho kinase inhibitor Y27632 to the key pathogenic cells in liver fibrosis, i.e. myofibroblasts including activated HSCs that highly express the PDGFβ-receptor, using the drug carrier pPB-MSA. This carrier consists of mouse serum albumin (MSA) covalently coupled to several PDGFβR-recognizing moieties (pPB). We aimed to create a prolonged release system of such a targeted construct, by encapsulating pPB-MSA-Y27632 in biodegradable polymeric microspheres, thereby reducing short-lasting peak concentrations and the need for frequent administrations. Firstly, we confirmed the vasodilating potency of PDGFβ-receptor targeted Y27632 in vitro in a contraction assay using HSCs seeded on a collagen gel. We subsequently demonstrated the in vivo antifibrotic efficacy of pPB-MSA-Y27632-loaded microspheres in the Mdr2-/- mouse model of progressive biliary liver fibrosis. A single subcutaneous microsphere administration followed by organ harvest one week later clearly attenuated liver fibrosis progression and significantly suppressed the expression of fibrosis related genes, such as several collagens, profibrotic cytokines and matrix metalloproteinases. In conclusion, we demonstrate that polymeric microspheres are suitable as drug delivery system for the sustained systemic delivery of targeted protein constructs with antifibrotic potential, such as pPB-MSA-Y27632. This formulation appears suitable for the sustained treatment of liver fibrosis and possibly other chronic diseases.

Keywords: Biologicals; Controlled release; Drug targeting; Liver fibrosis; Polymeric microspheres; Protein delivery.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

ATP Binding Cassette Transporter, Subfamily B / genetics
ATP-Binding Cassette Sub-Family B Member 4
Amides / administration & dosage*
Animals
Cell Line
Delayed-Action Preparations / administration & dosage
Drug Carriers / administration & dosage*
Female
Humans
Liver Cirrhosis / drug therapy*
Liver Cirrhosis / metabolism
Mice, Knockout
Microspheres
Protein Kinase Inhibitors / administration & dosage*
Pyridines / administration & dosage*
Receptor, Platelet-Derived Growth Factor beta / metabolism*
rho-Associated Kinases / antagonists & inhibitors*

Substances

ATP Binding Cassette Transporter, Subfamily B
Amides
Delayed-Action Preparations
Drug Carriers
Protein Kinase Inhibitors
Pyridines
Y 27632
Receptor, Platelet-Derived Growth Factor beta
rho-Associated Kinases